桂枝通络片通过抑制中性粒细胞胞外诱捕网缓解动脉粥样硬化  

Laxation Atherosclerosis of Guizhi Tongluo Tablets by Inhibiting Neutrophil Extracellular Trapping Nets

作　　者：吕秋云 杨林 沈东 黄小靖 潘显梅 钟森杰 陈洁[3] 王陵军[3,4] 冼绍祥 徐文华[5] 方红城 LYU Qiuyun;YANG Lin;SHEN Dong;HUANG Xiaojing;PAN Xianmei;ZHONG Senjie;CHEN Jie;WANG Lingjun;XIAN Shaoxiang;XU Wenhua;FANG Hongcheng(Graduate Training Base of Shenzhen Integrated Traditional Chinese and Western Medicine Hospital,Guangzhou University of Traditional Chinese Medicine,Shenzhen 518104,China;Department of Cardiovascular,Shenzhen Hospital of Integrated Chinese and Western Medicine,Shenzhen 518104,China;Department of Cardiovascular,the First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405,China;National Key Laboratory of Chinese Syndromes,Guangzhou 510405,China;Department of Preventive Treatment,Shenzhen Hospital of Integrated Chinese and Western Medicine,Shenzhen 518104,China)

机构地区：[1]广州中医药大学深圳市中西医结合医院研究生培养基地,深圳518104 [2]深圳市中西结合医院心血管科,深圳518104 [3]广州中医药大学第一附属医院心血管科,广州510405 [4]中国证候全国重点实验室,广州510405 [5]深圳市中西结合医院治未病科,深圳518104

出　　处：《医药导报》2024年第12期1898-1903,共6页Herald of Medicine

基　　金：深圳市“医疗卫生三名工程”(SZZYSM202106006);中国博士后科学基金资助项目(2023M730812);国家自然科学基金资助项目(82174316)。

摘　　要：目的探讨桂枝通络片(GZTLP)通过调控中性粒细胞胞外诱捕网(NETs)改善APOE基因敲除小鼠动脉粥样硬化的作用机制。方法将24只8周龄APOE基因敲除小鼠造模后随机分为4组:GZTLP大剂量组、小剂量组、模型对照组,正常对照组,每组各6只。GZTLP大剂量组、小剂量组分别给予GZTLP 1.87、0.47 mg·g-1灌胃,正常对照组和模型对照组灌胃给予等体积0.9%氯化钠溶液,灌胃6周,采用大体油红O染色整体观察主动脉内脂质斑块沉积情况;油红O染色观察主动脉根部脂质沉积;苏木精-伊红(HE)染色观察主动脉根部脂质斑块病理形态学改变;酶联免疫吸附测定(ELISA)检测小鼠外周血白细胞介素-1β(IL-1β)、肿瘤坏死因子α(TNF-α)水平;采用免疫荧光法检测主动脉弓部斑块部位淋巴细胞抗原6G(Ly6G)、中性粒细胞外诱捕网形成标志基因髓过氧化物酶(MPO)和瓜氨酸化组蛋白H3(Cit-H3)的表达及Ly6G、MPO和Cit-H3的共定位情况。结果与正常对照组相比,模型对照组小鼠主动脉出现严重的脂质斑块沉积,形态学损伤,大量炎症细胞浸润,血清炎症因子IL-1β和TNF-α的含量增多,主动脉组织中Ly6G、Cit-H3和MPO的蛋白表达明显增多;与模型对照组相比,GZTLP组小鼠主动脉内脂质斑块沉积斑块减少,主动脉细胞排列较规整,炎症细胞浸润情况改善,血清炎症因子IL-1β和TNF-α的含量显著减少(P<0.05),主动脉组织中Ly6G、MPO和Cit-H3发生共定位及蛋白表达明显减少(P<0.01)。结论GZTLP能改善动脉粥样硬化,其机制可能与抑制中性粒细胞胞外诱捕网有关。Objective To investigate the mechanism of Guizhi Tongluo Tablets(GZTLP)on improving atherosclerosis in APOE knockout mice by regulating neutrophil extracellular trapping nets(NETs).Methods After modeling,24 APOE knockout mice aged 8 weeks were randomly divided into 4 groups:GZTLP high-dose group,low-dose group,model control group and normal control group,with 6 mice in each group.GZTLP was given 1.87 mg·g-1 and 0.47 mg·g-1 intragastric administration in high-dose group and low-dose group,respectively.The normal control group and model control group were given 0.9%sodium chloride solution intragastric administration for 6 weeks,and the lipid plaque deposition in aorta was observed by gross oil red O staining.Lipid deposition in aortic root was observed by oil red O staining.The pathological changes of lipid plaques in aortic root were observed by HE staining.The levels of interleukin-1β(IL-1β)and tumor necrosis factorα(TNF-α)in peripheral blood of mice were detected by enzyme-linked immunosorbent assay(ELISA).The expression of lymphocyte antigen 6G(Ly6G),myeloperoxidase(MPO)and citrulinated histone(Cit-H3)in plaques of the aortic arch and the colocalization of Ly6G,MPO and Cit-H3 were detected by immunofluorescence assay.Results Compared with the normal control group,the aorta of mice in the model control group showed serious lipid plaque deposition,morphological damage,and a large number of inflammatory cells infiltration,the contents of serum inflammatory factors IL-1βand TNF-αwere increased,and the protein expressions of Ly6G,Cit-H3 and MPO were significantly increased.Compared with model control group,GZTLP group reduced the amount of lipid plaque deposition in aorta,the arrangement of aortic cells was more regular,the inflammatory cell infiltration was improved,and the contents of serum inflammatory factors IL-1βand TNF-αwere significantly decreased(P<0.05).The colocalization and the protein expression of Ly6G,MPO and Cit-H3 were significantly decreased in aortic tissues(P<0.01).Conclusions GZTLP ca

关 键 词：桂枝通络片 动脉粥样硬化 中性粒细胞 中性粒细胞外诱捕网 

分 类 号：R972.6[医药卫生—药品] R543[医药卫生—药学]